IP Library Granted Patent US 9,896,714
Granted Patent B2
US 9,896,714 · App. 13/518,785 · Granted Feb 20, 2018

Method of detecting phytase activity or protease activity

Inventor: Shukun Yu (Malmo, SE)
Assignee: DUPONT NUTRITION BIOSCIENCES APS
C12Q1/37C12Q1/42C12Q1/68G01N33/491G01N33/56966G01N33/57484
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Quick Facts
Patent No.
US 9,896,714
App. No.
13/518,785
Granted
Feb 20, 2018
Kind
B2
Abstract

A method of detecting a phytase activity or a protease activity is described. The method comprises the steps of: (a) providing a composition comprising a phytate/protein complex in a liquid or a gel; wherein the phytate/protein complex provides a detectable property to the composition; (b) providing a sample that comprises or is suspected of comprising phytase activity and/or protease activity, wherein the phytase and/or protease activity is capable of causing a change in the detectable property of the composition; (c) contacting the composition with the sample; and (d) determining if there is a detectable change in detectable property of the composition.

Claims (25)

1. A method of detecting a phytase activity or a protease activity comprising the steps of:

providing a medium comprising a continuous phase and a disperse phase, wherein the disperse phase comprises:

(1) a first component, the first component being a phytate or a phytic acid, and

(2) a second component, the second component being a protein,

wherein the first component and the second component are held together by one or more intermolecular interactions, wherein the phase provides a detectable property of turbidity to the medium;

providing a sample that comprises one or more of the phytase activity or the protease activity, wherein the one or more of the phytase activity or the protease activity are capable of affecting the disperse phase to cause a change in the turbidity of the medium;

contacting the medium with the sample; and

determining that there is a detectable change in the turbidity of the medium to determine an amount of the phytase activity or the protease activity in the sample.

2. The method of claim 1 , wherein the protein in the disperse phase of the medium is selected from a group comprising caseins, soy proteins, rapeseed proteins, mustard proteins, porcine haemoglobins, N,N-dimethylated caseins, bovine beta-lactoglobulins, bovine serum albumins, swine serum albumins, lysozymes, or proteins modified with a chromophore.

3. The method of claim 1 , wherein the medium is between a pH of 1.8 and a pH of 8.5.

4. The method of claim 1 , wherein the method comprises comparing the change in the detectable property of the medium to a control.

5. The method of claim 1 , wherein the sample is a biological sample.

6. The method of claim 5 , wherein the biological sample is an enzyme preparation comprised of a fermentation broth, a feed or in an extract of the fermentation broth or the feed.

7. The method of claim 1 , wherein determining if there is a detectable change in the detectable property of the medium comprises one or more of a quantitative assay, a semi-quantitative assay or a qualitative assay.

8. A method of detecting a phytase activity comprising the steps of:

providing a medium comprising a continuous phase and a disperse phase, wherein the disperse phase comprises:

(1) a first component, the first component being a phytate or a phytic acid, and

(2) a second component, the second component being a protein, wherein the first component and the second component are held together by one or more intermolecular interactions, wherein the disperse phase provides a detectable property of turbidity to the medium;

providing a sample that comprises the phytase activity, wherein the phytase activity is capable of affecting the disperse phase to cause a change in the turbidity of the medium;

contacting the medium with the sample, wherein one or more protease inhibitors are added to the sample in a sufficient amount as to inhibit any protease activity; and

determining that there is a detectable change in the turbidity of the medium to determine an amount of the phytase activity in the sample.

9. The method of claim 8 , wherein the protein in the disperse phase of the medium is selected from a group comprising caseins, soy proteins, rapeseed proteins, mustard proteins, porcine haemoglobins, N,N-dimethylated caseins, bovine beta-lactoglobulins, bovine serum albumins, swine serum albumins, lysozymes, or proteins modified with a chromophore.

10. The method of claim 8 , wherein the sample is a biological sample.

11. The method of claim 10 , wherein the biological sample is an enzyme preparation comprised of a fermentation broth, a feed or in an extract of the fermentation broth or the feed.

12. The method of claim 8 , wherein determining if there is a detectable change in the detectable property of the medium comprises one or more of a quantitative assay, a semi-quantitative assay or a qualitative assay.

Assignments (2)
CHANGE OF NAME Recorded Sep 10, 2012
From: DANISCO A/S
To: DUPONT NUTRITION BIOSCIENCES APS
Reel/Frame 028930/0898 →
ASSIGNMENT OF ASSIGNOR'S INTEREST Recorded Sep 7, 2012
From: YU, SHUKUN
To: DANISCO A/S
Reel/Frame 028914/0931 →
Priority Claims (1)
EP 10172171 · Aug 6, 2010 · regional
Continuity (2)
Provisional Application 61298670 · Jan 27, 2010
Related Publication 20120264153A1 · Oct 18, 2012